Figure 5. ADAM17 expressed on MCF-7 dose dependently degrades recombinant IFN-γ.

(A,B) Real-time PCR analysis of ADAM17 mRNA expression in overexpressed or knocked down MCF-7. Fold changes from control were shown. *p < 0.05 versus control. ©Protein level expression of ADAM17 on the cell surface of MCF-7. Fc receptor blocking solution (Biolegend, San Diego, CA) was used prior to antibody application. The results of flow cytometry analysis are shown. Black dotted line indicates the result of fluorophore-conjugated isotype IgG. Blue, red, and green lines indicate the result of control cells, overexpressed cells, and knocked down cells, respectively. (D) Mean percent of positive cells from the results of flowcytometry analysis are shown. *p < 0.05 versus control. (E) IFN-γ concentration measured by ELISA. 1 ng/mL of recombinant mouse IFN-γ were added into each culture media, and incubated for 1 hour. *p < 0.05 versus control. (F) Western blot analysis of IFN-γ performed using the collected culture supernatant. tf: overexpressed cells. RNAi: knocked down cells. L: molecular weight marker.